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Effects on fertility

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Route of administration:
oral: gavage
Vehicle:
polyethylene glycol
Remarks:
PEG 400 was dissolved in distilled water to be the ratio of 80% (v/v).
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Samples for homogeneity analysis were also analyzed for verification of dose level concentration. Results of dose formulation were 94.8, 91.1 and 104.5% at each dose levels of 25, 75 and 250 mg/mL. They were acceptable as the mean concentration was within ± 20% of the nominal concentration.
Duration of treatment / exposure:
Dosing of the males will begin 14 days prior to mating and continue through the day prior to sacrifice (at least 28 days). Dosing of the females will begin 14 days prior to mating and continue through lactation day (LD) 13. Animals in recovery group will not be mated and will be assigned to 2 weeks of recovery period after the completion of administration
Frequency of treatment:
once a day
Dose / conc.:
0 mg/kg bw (total dose)
Dose / conc.:
100 mg/kg bw (total dose)
Dose / conc.:
300 mg/kg bw (total dose)
Dose / conc.:
1 000 mg/kg bw (total dose)
No. of animals per sex per dose:
control: 18 animals
100, 300 mg/kg: 12 animals
1000 mg/kg: 18 animals
Control animals:
yes
Parental animals: Observations and examinations:
One male to one female mating was used, and males and females were paired for 2 weeks. Animals of the recovery group were not mated.
Mating confirmation was checked every morning during the mating period. Mating was confirmed with vaginal plug(s) and/or sperm in the vaginal smear. Day 0 of pregnancy was defined as the day of mating confirmation. Pregnancy was confirmed by parturition or by implantation sites on the uterus at sacrifice.
Non-mated female was sacrificed after at least 24 days from final mating day as described in
Section 2.13.1, and dosing was continued until the day before of sacrifice.
Oestrous cyclicity (parental animals):
A vaginal smear was taken daily for each female from the beginning of the 14 days prior to mating with continued monitoring into the mating period until there was evidence of mating.
Furthermore, regularity and length of the estrus cycle during the treatment period until mating was examined.
In addition, vaginal smear of sacrificed females were taken at termination to examine the stage of the estrus cycle and to allow a correlation with the histopathology of the female reproductive organs.
Postmortem examinations (offspring):
External and visceral examinations were conducted in dead pups at parturition and in dead or moribund pups from birth to day 4 of lactation, if possible. After day 5 of lactation, dead pups were sacrificed by CO2 inhalation and necropsied with special attention to all vital organs.
Reproductive indices:
• Mating Index (%)
= (No. of males with evidence of mating/No. of males paired) × 100
= (No. of females with evidence of mating/No. of females paired) × 100

• Fertility Index (%)
= (No. of males impregnating a female/No. of males paired) × 100
= (No. of pregnant females/No. of females paired) × 100

• Fecundity Index and Pregnancy Index (%)
= (No. of males impregnating a female/No. of males with evidence of mating) × 100
= (No. of pregnant females/No. of females with evidence of mating) × 100

• Precoital Time: No. of days taken to mate
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
In males at 100, 300 and 1000 mg/kg, salivation was observed in 11, 12, and 18 animals, respectively. In females at 100, 300 and 1000 mg/kg, salivation was observed in 9, 11, and 18 animals, respectively. It was considered test item-related but not toxicologically statistically significant since it was considered to be attributed to the palatability of the test item.
Dermal irritation (if dermal study):
not examined
Mortality:
mortality observed, non-treatment-related
Description (incidence):
No test item-related deaths or moribund animals occurred in any group throughout the study.
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
No test item-related changes in body weight and body weight gain were observed in both sexes during the study.
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
No test item-related changes in food consumption were observed in both sexes during the study.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
Total red blood cell count (RBC) was significantly increased (1.07-fold over control, respectively) in males at 300 and 1000 mg/kg and mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH) were significantly increased (1.08 and 1.07-fold over control,
respectively) in females at 1000 mg/kg. These changes were fully recovered after recovery period and considered not to represent meaningful toxicity as there were no histopathological correlates.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
Inorganic phosphorus (IP) was significantly decreased (85% of control) in males at 1000 mg/kg. These changes were fully recovered after recovery period and considered not to represent meaningful toxicity as there were no histopathological correlates.
Urinalysis findings:
not examined
Behaviour (functional findings):
effects observed, non-treatment-related
Description (incidence and severity):
No test item-related changes in functional behavior examination were observed in both sexes during the study.
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
No test item-related changes in organ weights were observed in both sexes during the study.
Gross pathological findings:
not examined
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Reproductive function: oestrous cycle:
effects observed, non-treatment-related
Description (incidence and severity):
No test item-related changes in estrus cycle were observed during the study.
Reproductive function: sperm measures:
not specified
Reproductive performance:
effects observed, non-treatment-related
Description (incidence and severity):
No test item-related changes in the reproductive and littering findings were observed during the study.
Key result
Dose descriptor:
NOAEL
Effect level:
<= 1 000 mg/kg bw (total dose)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
reproductive function (oestrous cycle)
reproductive performance
Key result
Critical effects observed:
no
Lowest effective dose / conc.:
1 000 mg/kg bw (total dose)
Clinical signs:
not examined
Dermal irritation (if dermal study):
not examined
Mortality:
not examined
Body weight and weight changes:
not examined
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
not examined
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
not examined
Reproductive performance:
not examined
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
No test item-related changes in clinical signs were observed in both sexes of the F1 pups during the study.
Dermal irritation (if dermal study):
not examined
Mortality / viability:
mortality observed, non-treatment-related
Description (incidence and severity):
No test item-related changes in the external examination were observed in both sexes of the F1 culled pups during the study.
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
No test item-related changes in body weight were observed in both sexes of the F1 pups during the study.
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
not examined
Histopathological findings:
not examined
Other effects:
effects observed, non-treatment-related
Description (incidence and severity):
1. F1 pups anogenital distance
No test item-related changes in anogenital distance (AGD) were observed in both sexes of F1 pups during the study.
Statistically significant changes in F1 male pups AGD was not considered test item-related since it was within the historical control data. [Normalized AGD (Min-Max, mm); Male: 2.0-2.2]

2. F1 male pups nipple retention
No test item-related changes in nipple retention of the F1 male pups were observed during the study.
Behaviour (functional findings):
effects observed, non-treatment-related
Description (incidence and severity):
Thyroid Hormone (T4) analysis:
No test item-related changes in the thyroid hormone (T4) were observed in the adult male animals, and in the male and female pups on PND 13.
Developmental immunotoxicity:
not examined
Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
<= 1 000 mg/kg bw (total dose)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
clinical signs
mortality
body weight and weight gain
developmental neurotoxicity
Key result
Critical effects observed:
no
Lowest effective dose / conc.:
1 000 mg/kg bw (total dose)
Clinical signs:
not examined
Dermal irritation (if dermal study):
not examined
Mortality / viability:
not examined
Body weight and weight changes:
not examined
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
not examined
Histopathological findings:
not examined
Other effects:
not examined
Behaviour (functional findings):
not examined
Developmental immunotoxicity:
not examined
Key result
Reproductive effects observed:
no
Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available

Effects on developmental toxicity

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available

Justification for classification or non-classification